1. Tunable control of CAR T cell activity through tetracycline mediated disruption of protein–protein interaction.
- Author
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Hotblack, Alastair, Kokalaki, Evangelia K., Palton, Morgan J., Cheung, Gordon Weng-Kit, Williams, Iwan P., Manzoor, Somayya, Grothier, Thomas I., Piapi, Alice, Fiaccadori, Valeria, Wawrzyniecka, Patrycja, Roddy, Harriet A., Agliardi, Giulia, Roddie, Claire, Onuoha, Shimobi, Thomas, Simon, Cordoba, Shaun, and Pule, Martin
- Subjects
T cells ,PROTEIN-protein interactions ,AUTOMOBILES ,TETRACYCLINES ,TETRACYCLINE ,AMINO acid sequence - Abstract
Chimeric antigen receptor (CAR) T cells are a promising form of cancer immunotherapy, although they are often associated with severe toxicities. Here, we present a split-CAR design incorporating separate antigen recognition and intracellular signaling domains. These exploit the binding between the tetracycline repressor protein and a small peptide sequence (TIP) to spontaneously assemble as a functional CAR. Addition of the FDA-approved, small molecule antibiotic minocycline, acts as an "off-switch" by displacing the signaling domain and down-tuning CAR T activity. Here we describe the optimization of this split-CAR approach to generate a CAR in which cytotoxicity, cytokine secretion and proliferation can be inhibited in a dose-dependent and reversible manner. Inhibition is effective during on-going CAR T cell activation and inhibits activation and tumor control in vivo. This work shows how optimization of split-CAR structure affects function and adds a novel design allowing easy CAR inhibition through an FDA-approved small molecule. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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